sepharose and tricalcium-phosphate

Developing a simple produces for efficient derivation of motor neurons (MNs) is essential for neural tissue engineering studies. Stem cells with high capacity for neural differentiation and scaffolds with the potential to promote motor neurons differentiation are promising candidates for neural tissue engineering. Recently, human olfactory ecto-mesenchymal stem cells (OE-MSCs), which are isolated easily from the olfactory mucosa, are considered a new hope for neuronal replacement due to their neural crest origin. Herein, we synthesized conducting hydrogels using different concentration of chitosan-g-aniline pentamer, gelatin, and agarose. The chemical structures, swelling and deswelling ratio, ionic conductivity and thermal properties of the hydrogel were characterized. Scaffolds with 10% chitosan-g-aniline pentamer/gelatin (S10) were chosen for further investigation and the potential of OE-MSCs as a new source for programming to motor neuron-like cells investigated on tissue culture plate (TCP) and conductive hydrogels. Cell differentiation was evaluated at the level of mRNA and protein synthesis and indicated that conductive hydrogels significantly increased the markers related to motor neurons including Hb-9, Islet-1 and ChAT compared to TCP. Taken together, the results suggest that OE-MSCs would be successfully differentiated into motor neuron-like cells on conductive hydrogels and would have a promising potential for treating motor neuron-related diseases.

Topics: Aniline Compounds; Calcium Phosphates; Cell Differentiation; Cell Proliferation; Cell Survival; Cells, Cultured; Chitosan; Compressive Strength; Electric Conductivity; Gelatin; Humans; Hydrogels; Male; Mesenchymal Stem Cells; Motor Neurons; Olfactory Bulb; RNA, Messenger; Sepharose; Spectroscopy, Fourier Transform Infrared; Temperature; Thermogravimetry; Tissue Scaffolds

In this work a multifaceted approach to the fabrication of scaffolds is considered, that is, besides the preparation technique, the introduction of substances that may contribute to enhance their final performance, as well as the techniques required to ensure the correct preservation of the so obtained scaffolds are taken into account to tailor the release of vancomycin from beta-tricalcium phosphate (beta-TCP)/agarose scaffolds. These materials were prepared by a shaping technique that allows to obtain pieces at a temperature low enough to simultaneously include active substances susceptible of heat degrading such as vancomycin, the model drug considered in this work. In the first approximation poly(ethylene glycol) (PEG), a hydrophilic substance employed as a matrix capable of binding compounds such as proteins or peptides and release them in a controlled fashion, was included in the formulation. The second tool to govern the vancomycin liberation is based on the drying procedures employed to process and preserve the obtained scaffolds: freeze-drying and heat desiccation at 37 degrees C. These modifications resulted in the generation of different pore architectures and certain chemical interactions, such as the formation of an agarose-PEG-vancomycin complex that yielded different drug release patterns. The so obtained pieces behave like a hydrogel when immersed in a hydrated medium but show a consistency comparable to that of the cancellous bone.

Topics: Anti-Bacterial Agents; Calcium Phosphates; Chemistry, Pharmaceutical; Delayed-Action Preparations; Desiccation; Drug Carriers; Drug Compounding; Freeze Drying; Microscopy, Electron, Scanning; Porosity; Sepharose; Solubility; Temperature; Vancomycin; X-Ray Diffraction